SBIR Phase II: High-throughput Small RNA Sequencing

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is the development of a technology to accurately measure small RNA expression. This is an enabling life science research tool. Small RNAs are ubiquitous gene regulators found in the body. Products of the same microRNA gene that vary in length by one or two nucleotides may be involved in a host of diseases, including cancer. The value for developing a method to measure the true profile of microRNAs in a sample would be immense for the research community studying transcriptional regulation, and would open the doors to those interested in drug development and diagnostics. The goal of this proposal is to develop a library preparation kit for non-biased small RNA libraries for Next Generation sequencing. These kits will increase the quality and rate at which global microRNA profiles may be determined for research and clinical applications. This SBIR Phase II project proposes to develop next generation sequencing technology for small RNA more quantitative and less biased. High throughput sequencing has transformed the landscape of genomic research with its ability to produce gigabases of data in a single run. This has enabled researchers to perform genome wide and high depth sequencing studies that would normally not be possible. Despite this capacity, amplification artifacts introduced during polymerase chain reaction (PCR) assays increase the chance of duplicate reads and uneven distribution of read coverage. Accurate profiling using deep sequencing also has been undermined by biases with over- or under-represented microRNAs. The presence of these biases significantly limits the incredible sensitivity and accuracy made possible by next generation sequencing. The goal of this proposal is to develop novel bias-reducing technology for making small RNA libraries. The proposed kits and protocols will increase the rate at which global microRNA profiles can be determined, and between-sample and within-sample differences (as well as newly discovered small RNAs) can be subsequently validated. This product will result in a major shift in the way small RNA sequencing is performed, and will pave the way for the discovery of new small RNAs.